摘要
黄龙病是柑橘生产上的一种毁灭性病害,由韧皮部限制性细菌Candidatus Liberibacter导致。感染黄龙病的柑橘树的叶片有异常高淀粉积累,因此开发出了基于碘-淀粉显色法的黄龙病快速检测试剂盒。将不同浓度的可溶性淀粉溶液进行碘-淀粉显色反应,制作成快速检测试剂盒配套的比色卡,用来快速确认叶片淀粉值区间。根据不同采样时间的健康树和黄龙病树的叶片淀粉值差异进行统计数据分析,确定了7—10月为黄龙病快速检测试剂盒的适宜采样检测时间。根据显色易辨性和检测准确率将淀粉质量浓度3000 ppm确定为适宜的黄龙病阳性叶片淀粉值界限,即检测叶片淀粉值≥3000 ppm判定为黄龙病阳性。通过田间随机取样发现,快速检测试剂盒与实时荧光定量PCR检测方法的一致性约为80%。该黄龙病快速检测试剂盒对田间黄龙病的快速筛查具有重要意义,可作为实时荧光定量PCR检测方法的前序筛查辅助手段。
Huanglongbing is a destructive disease of citrus caused by phloem-limited bacteria,Candidatus Liberibacter spp.Leaves of citrus trees infected by Huanglongbing accumulate abnormal high starch content.Therefore,a rapid detection kit for huanglongbing is developed by an iodine-starch chromogenic reaction.Different concentrations of soluble starch solution are used for iodine-starch color development reaction,and a colorimetric card supporting the rapid detection kit is made to quickly confirm the starch content value range of leaves.Starch content values of leaves of healthy citrus trees are analyzed and compared with those of citrus trees infected by Huanglongbing in different sampling time on field test.It is found that suitable sampling time for huanglongbing rapid detection kit is from July to October.According to the color rendering and easy identification,the starch mass concentration of 3000 ppm is determined as the appropriate starch content value limit of Huanglongbing positive leaves,that is,when the starch content value of the leaf is detected≥3000 ppm,it is judged to be positive for Huanglongbing.Consistent rate between huanglongbing rapid detection kit method and real-time fluorescent quantitative PCR method is about 80%,which is further confirmed by random field sampling test.This Huanglongbing rapid detection kit is of high significance for field rapid screening of Huanglongbing disease,it can be used as an auxiliary means of pre-screening of real-time PCR detection methods.
作者
邓方坤
蔡德珍
欧阳智刚
翁雅青
王希
吴晓晖
刘新军
胡珊玲
陈红丽
DENG Fangkun;CAI Dezhen;OUYANG Zhigang;WENG Yaqing;WANG Xi;WU Xiaohui;LIU Xinjun;HU shanling;CHEN Hongli(Jiangxi Academy of Science,Institute of Applied Chemistry,Nanchang 330096,China;Jiangxi New Dragon Biotechnology Co.,Ltd.,Yichun 336000,China;School of Life Science,Gannan Normal University,Ganzhou 341000,China;Jiangxi Agricultural and Rural Development Service Center,Nanchang 330299,China;Dayu Bureau of Agriculture and Rural Affairs,Dayu 341500,China)
出处
《生物化工》
2023年第2期102-108,113,共8页
Biological Chemical Engineering
基金
江西省重大科技研发专项(20194ABC28007)
江西省科学院省级科研项目经费包干制试点示范项目(2022YSBG21022)。
关键词
柑橘黄龙病
快速检测试剂盒
碘-淀粉法
比色卡
实时荧光定量PCR
Citrus Huanglongbing
rapid detection kit
iodine-starch chromogenic reaction
colourimetric card
real-time fluorescent quantitative PCR
